NS5A regulates PC expression level.

<p>(A–C) PC expression level is decreased, whereas FAS expression level is increased in cells expressing NS5A protein. (A) Total cell lysates harvested from either IFN-cured cells or replicon cells were immunoblotted with the indicated antibodies (top panel). Total RNAs isolated from either IFN-cured cells or replicon cells were quantitated for PC mRNA levels (middle panel) and FAS mRNA levels (bottom panel) by qRT-PCR. (B) Huh7.5 cells were either mock infected or infected with Jc1. Total cell lysates harvested at 5 days postinfection were immunoblotted with the indicated antibodies (top panel). Total RNAs isolated from either mock infected or Jc1 infected cells were quantitated for PC mRNA levels (middle panel) and FAS mRNA levels (bottom panel) by qRT-PCR using data from three independent experiments. Asterisks indicate significant differences (*, <i>P</i><0.05) from the value for the control. Error bars indicate standard deviations. (C) Equal amounts of cell lysates harvested from either vector stable or NS5A stable cells derived from genotype 1b or 2a were immunoblotted with the indicated antibodies (top panel). Total RNAs isolated from the indicated stable cells were quantitated for PC mRNA levels (middle panel) and FAS mRNA levels (bottom panel). Asterisks indicate significant differences (*, <i>P</i><0.05) from the value for the control. (D) Huh7.5 cells were transfected with siRNAs of either negative or PC. At 3 days after transfection, cells were harvested and analyzed by immunoblotting with the indicated antibodies (top panel). Actin was used as a loading control. Total RNAs isolated from the indicated cells were quantitated for PC mRNA levels (middle panel) and FAS mRNA levels (bottom panel). Asterisks indicate significant differences (*, <i>P</i><0.05) from the value for the control. (E) Huh7.5 cells were transiently transfected with either pEF6 empty vector or pEF6-NS5A-Myc expression plasmid. Cell lysates harvested at 48 h or 72 h after transfection were immunoblotted with the indicated antibodies.